Tandem chemical deconstruction and biological upcycling of poly(ethylene terephthalate) to β-ketoadipic acid by Pseudomonas putida KT2440

Allison Z. Werner; Rita Clare; Thomas D. Mand; Isabel Pardo; Kelsey J. Ramirez; Stefan J. Haugen; Felicia Bratti; Gara N. Dexter; Joshua R. Elmore; Jay D. Huenemann; George L. Peabody; Christopher W. Johnson; Nicholas A. Rorrer; Davinia Salvachúa; Adam M. Guss; Gregg T. Beckham

doi:10.1016/j.ymben.2021.07.005

Tandem chemical deconstruction and biological upcycling of poly(ethylene terephthalate) to β-ketoadipic acid by Pseudomonas putida KT2440

Allison Z. Werner, Rita Clare, Thomas D. Mand, Isabel Pardo, Kelsey J. Ramirez, Stefan J. Haugen, Felicia Bratti, Gara N. Dexter, Joshua R. Elmore, Jay D. Huenemann, George L. Peabody, Christopher W. Johnson, Nicholas A. Rorrer, Davinia Salvachúa, Adam M. Guss, Gregg T. Beckham

Synthetic Biology

Research output: Contribution to journal › Article › peer-review

95 Scopus citations

Abstract

Poly(ethylene terephthalate) (PET) is the most abundantly consumed synthetic polyester and accordingly a major source of plastic waste. The development of chemocatalytic approaches for PET depolymerization to monomers offers new options for open-loop upcycling of PET, which can leverage biological transformations to higher-value products. To that end, here we perform four sequential metabolic engineering efforts in Pseudomonas putida KT2440 to enable the conversion of PET glycolysis products via: (i) ethylene glycol utilization by constitutive expression of native genes, (ii) terephthalate (TPA) catabolism by expression of tphA2_IIA3_IIB_IIA1_II from Comamonas and tpaK from Rhodococcus jostii, (iii) bis(2-hydroxyethyl) terephthalate (BHET) hydrolysis to TPA by expression of PETase and MHETase from Ideonella sakaiensis, and (iv) BHET conversion to a performance-advantaged bioproduct, β-ketoadipic acid (βKA) by deletion of pcaIJ. Using this strain, we demonstrate production of 15.1 g/L βKA from BHET at 76% molar yield in bioreactors and conversion of catalytically depolymerized PET to βKA. Overall, this work highlights the potential of tandem catalytic deconstruction and biological conversion as a means to upcycle waste PET.

Original language	English
Pages (from-to)	250-261
Number of pages	12
Journal	Metabolic Engineering
Volume	67
DOIs	https://doi.org/10.1016/j.ymben.2021.07.005
State	Published - Sep 2021

Funding

This work was authored in part by the National Renewable Energy Laboratory, operated by Alliance for Sustainable Energy, LLC, for the U.S. Department of Energy (DOE) under Contract No. DE-AC36-08GO28308. This work was authored in part by Oak Ridge National Laboratory, which is managed by UT-Battelle, LLC, for the U.S. DOE under contract DE-AC05-00OR22725. Funding was provided by the U.S. Department of Energy, Office of Energy Efficiency and Renewable Energy, Advanced Manufacturing Office (AMO) and Bioenergy Technologies Office (BETO). This work was performed as part of the BOTTLE™ Consortium and was supported by AMO and BETO under contract no. DE-AC36-08GO28308 with the National Renewable Energy Laboratory, operated by Alliance for Sustainable Energy, LLC and contract no. DE-AC05-00OR22725 with Oak Ridge National Laboratory, operated by UT-Battelle. The views expressed in the article do not necessarily represent the views of the DOE or the U.S. Government. The U.S. Government retains and the publisher, by accepting the article for publication, acknowledges that the U.S. Government retains a nonexclusive, paid-up, irrevocable, worldwide license to publish or reproduce the published form of this work, or allow others to do so, for U.S. Government purposes. We thank McKenzie Hawkins, William E. Michener, and Joel Miscall for analytical support, Lahiru N. Jayakody for providing the pLJ062 plasmid, and Lindsay Eltis for kindly providing Rhodococcus jostii RHA1. This work was authored in part by the National Renewable Energy Laboratory, operated by Alliance for Sustainable Energy, LLC, for the U.S. Department of Energy (DOE) under Contract No. DE-AC36-08GO28308 . This work was authored in part by Oak Ridge National Laboratory, which is managed by UT-Battelle, LLC, for the U.S. DOE under contract DE-AC05-00OR22725 . Funding was provided by the U.S. Department of Energy, Office of Energy Efficiency and Renewable Energy , Advanced Manufacturing Office (AMO) and Bioenergy Technologies Office (BETO) . This work was performed as part of the BOTTLE™ Consortium and was supported by AMO and BETO under contract no. DE-AC36-08GO28308 with the National Renewable Energy Laboratory, operated by Alliance for Sustainable Energy, LLC and contract no. DE-AC05-00OR22725 with Oak Ridge National Laboratory, operated by UT-Battelle. The views expressed in the article do not necessarily represent the views of the DOE or the U.S. Government. The U.S. Government retains and the publisher, by accepting the article for publication, acknowledges that the U.S. Government retains a nonexclusive, paid-up, irrevocable, worldwide license to publish or reproduce the published form of this work, or allow others to do so, for U.S. Government purposes. We thank McKenzie Hawkins, William E. Michener, and Joel Miscall for analytical support, Lahiru N. Jayakody for providing the pLJ062 plasmid, and Lindsay Eltis for kindly providing Rhodococcus jostii RHA1.

Keywords

Bio-upcycling
MHETase
Metabolic engineering
PETase
Plastics upcycling
Terephthalic acid

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.ymben.2021.07.005

Cite this

Werner, A. Z., Clare, R., Mand, T. D., Pardo, I., Ramirez, K. J., Haugen, S. J., Bratti, F., Dexter, G. N., Elmore, J. R., Huenemann, J. D., Peabody, G. L., Johnson, C. W., Rorrer, N. A., Salvachúa, D., Guss, A. M., & Beckham, G. T. (2021). Tandem chemical deconstruction and biological upcycling of poly(ethylene terephthalate) to β-ketoadipic acid by Pseudomonas putida KT2440. Metabolic Engineering, 67, 250-261. https://doi.org/10.1016/j.ymben.2021.07.005

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title = "Tandem chemical deconstruction and biological upcycling of poly(ethylene terephthalate) to β-ketoadipic acid by Pseudomonas putida KT2440",

abstract = "Poly(ethylene terephthalate) (PET) is the most abundantly consumed synthetic polyester and accordingly a major source of plastic waste. The development of chemocatalytic approaches for PET depolymerization to monomers offers new options for open-loop upcycling of PET, which can leverage biological transformations to higher-value products. To that end, here we perform four sequential metabolic engineering efforts in Pseudomonas putida KT2440 to enable the conversion of PET glycolysis products via: (i) ethylene glycol utilization by constitutive expression of native genes, (ii) terephthalate (TPA) catabolism by expression of tphA2IIA3IIBIIA1II from Comamonas and tpaK from Rhodococcus jostii, (iii) bis(2-hydroxyethyl) terephthalate (BHET) hydrolysis to TPA by expression of PETase and MHETase from Ideonella sakaiensis, and (iv) BHET conversion to a performance-advantaged bioproduct, β-ketoadipic acid (βKA) by deletion of pcaIJ. Using this strain, we demonstrate production of 15.1 g/L βKA from BHET at 76% molar yield in bioreactors and conversion of catalytically depolymerized PET to βKA. Overall, this work highlights the potential of tandem catalytic deconstruction and biological conversion as a means to upcycle waste PET.",

keywords = "Bio-upcycling, MHETase, Metabolic engineering, PETase, Plastics upcycling, Terephthalic acid",

author = "Werner, {Allison Z.} and Rita Clare and Mand, {Thomas D.} and Isabel Pardo and Ramirez, {Kelsey J.} and Haugen, {Stefan J.} and Felicia Bratti and Dexter, {Gara N.} and Elmore, {Joshua R.} and Huenemann, {Jay D.} and Peabody, {George L.} and Johnson, {Christopher W.} and Rorrer, {Nicholas A.} and Davinia Salvach{\'u}a and Guss, {Adam M.} and Beckham, {Gregg T.}",

note = "Publisher Copyright: {\textcopyright} 2021 The Authors",

year = "2021",

month = sep,

doi = "10.1016/j.ymben.2021.07.005",

language = "English",

volume = "67",

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Werner, AZ, Clare, R, Mand, TD, Pardo, I, Ramirez, KJ, Haugen, SJ, Bratti, F, Dexter, GN, Elmore, JR, Huenemann, JD, Peabody, GL, Johnson, CW, Rorrer, NA, Salvachúa, D, Guss, AM & Beckham, GT 2021, 'Tandem chemical deconstruction and biological upcycling of poly(ethylene terephthalate) to β-ketoadipic acid by Pseudomonas putida KT2440', Metabolic Engineering, vol. 67, pp. 250-261. https://doi.org/10.1016/j.ymben.2021.07.005

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T1 - Tandem chemical deconstruction and biological upcycling of poly(ethylene terephthalate) to β-ketoadipic acid by Pseudomonas putida KT2440

AU - Werner, Allison Z.

AU - Clare, Rita

AU - Mand, Thomas D.

AU - Pardo, Isabel

AU - Ramirez, Kelsey J.

AU - Haugen, Stefan J.

AU - Bratti, Felicia

AU - Dexter, Gara N.

AU - Elmore, Joshua R.

AU - Huenemann, Jay D.

AU - Peabody, George L.

AU - Johnson, Christopher W.

AU - Rorrer, Nicholas A.

AU - Salvachúa, Davinia

AU - Guss, Adam M.

AU - Beckham, Gregg T.

PY - 2021/9

Y1 - 2021/9

N2 - Poly(ethylene terephthalate) (PET) is the most abundantly consumed synthetic polyester and accordingly a major source of plastic waste. The development of chemocatalytic approaches for PET depolymerization to monomers offers new options for open-loop upcycling of PET, which can leverage biological transformations to higher-value products. To that end, here we perform four sequential metabolic engineering efforts in Pseudomonas putida KT2440 to enable the conversion of PET glycolysis products via: (i) ethylene glycol utilization by constitutive expression of native genes, (ii) terephthalate (TPA) catabolism by expression of tphA2IIA3IIBIIA1II from Comamonas and tpaK from Rhodococcus jostii, (iii) bis(2-hydroxyethyl) terephthalate (BHET) hydrolysis to TPA by expression of PETase and MHETase from Ideonella sakaiensis, and (iv) BHET conversion to a performance-advantaged bioproduct, β-ketoadipic acid (βKA) by deletion of pcaIJ. Using this strain, we demonstrate production of 15.1 g/L βKA from BHET at 76% molar yield in bioreactors and conversion of catalytically depolymerized PET to βKA. Overall, this work highlights the potential of tandem catalytic deconstruction and biological conversion as a means to upcycle waste PET.

AB - Poly(ethylene terephthalate) (PET) is the most abundantly consumed synthetic polyester and accordingly a major source of plastic waste. The development of chemocatalytic approaches for PET depolymerization to monomers offers new options for open-loop upcycling of PET, which can leverage biological transformations to higher-value products. To that end, here we perform four sequential metabolic engineering efforts in Pseudomonas putida KT2440 to enable the conversion of PET glycolysis products via: (i) ethylene glycol utilization by constitutive expression of native genes, (ii) terephthalate (TPA) catabolism by expression of tphA2IIA3IIBIIA1II from Comamonas and tpaK from Rhodococcus jostii, (iii) bis(2-hydroxyethyl) terephthalate (BHET) hydrolysis to TPA by expression of PETase and MHETase from Ideonella sakaiensis, and (iv) BHET conversion to a performance-advantaged bioproduct, β-ketoadipic acid (βKA) by deletion of pcaIJ. Using this strain, we demonstrate production of 15.1 g/L βKA from BHET at 76% molar yield in bioreactors and conversion of catalytically depolymerized PET to βKA. Overall, this work highlights the potential of tandem catalytic deconstruction and biological conversion as a means to upcycle waste PET.

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KW - MHETase

KW - Metabolic engineering

KW - PETase

KW - Plastics upcycling

KW - Terephthalic acid

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SN - 1096-7176

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JF - Metabolic Engineering

ER -

Tandem chemical deconstruction and biological upcycling of poly(ethylene terephthalate) to β-ketoadipic acid by Pseudomonas putida KT2440

Abstract

Funding

Keywords

UN SDGs

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